Analysis Affinity of Liver Cancer Antigen Roundabout1 (Robo1) by KinExA (CAT#: STEM-MB-0017-CJ)

Introduction

Human ROBO1 contains five immunoglobulin domains, three fibronectin III domains, a transmembrane domain, and an intracellular tail. ROBO1 is a member of the neural cell adhesion molecule family of receptors. Meanwhile ROBO1 was cloned as Deleted in U Twenty Twenty (DUTT1) from the lung cancer tumor suppressor gene region at 3p12. ROBO1 mRNA expression is silenced in patients with lung and breast cancer caused by hemizygous and homozygous deletions and/or hypermethylation of the promoter region of the ROBO1 gene. Therefore, ROBO1 is thought to be a candidate tumor suppressor gene.

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Principle Applications Procedure Materials Notes Related Products

Principle

KinExA is a two-stage analysis system. In the first stage, a number of solutions are prepared, where one partner remains constant (constant binding partner, or CBP) and the other (titrant) is variable, usually serially diluted. As the titrant is added, the free CBP decreases and is analysed by a sophisticated and precise microfluorescence measurement device. The signal generated can be mathematically related to the affinity (KD) of the two molecules for each other, as well as the kinetic parameters of binding (kon) and dissociation (koff).

Applications

Oncology & Cancer; Immunology/Inflammation; Pharmacology

Procedure

1. preparation of the functionalized beads which will capture the analyte for measuremen.
2. preparation of a series of solutions consisting of a constant initial concentration of one component of the binary reaction and serial dilutions of the other reactant. The component that is kept constant is the constant binding partner (CBP) , and is the one which will be analyzed.
3. each reaction mixture is sampled and the fluorescence of free CBP bound to the capture beads is obtained for subsequent numerical analysis.

Materials

• Sample Type: Serum, Plasma, Urine
• Equipment: Kinetic Exclusion Assay (KinExA)

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